Experimental Study On The Effects Of The Bushen Huatan Fang On Osteogenesis In Osteoporosis Rats And Its Possible Mechanisms

Objective:This research subject is plans to treat the ovariectomized rats with the traditional Chinese medicine compound prescription of the Bushen Hutan Therapy and Li vial, and be observed ovary osteoporosis model rats the bone mineral density,N-midBGP,25 (OH) D and PICP in the blood serum, BGPmRNA and CbfalmRNA expression in bone marrow cells, BMP-2 and TGF-β1 expression in bone tissues to discuss the effection of the Bushen Hutan Fang on osteoporosis in ovariectomized rats and its hossible mechanisms.Methods:We made rat osteoporosis model by ablating ovary in the experiment. The experiment rat were divided to five groups:the sham operation group,the model group,the western medicine group,the nomal dose herb group of Bushen Hutan Therapy and the much dose herb group of Bushen Hutan Therapy. The treatment began three months after the surgery and lasted for two mouths. The western medicine group used teblet of Livial as contrast. The rats in sham operation group and in model group were administered by 0.9%Nacl liquid. All rats from each experimentation groups had been sacrificed separately at two months after the medication. We had used dual energy X-ray bone density instrument to detect the bone mineral density of the rats; detected the activity of N-MID,25 (OH) VD and PICP in the blood serum of the rat swith the method of ELISA. We had used the RT-PCR method to investigate the expression of BGPmRNA and CbfalmRNA in bone marrow cells, observed the morpholoy of the bone tissues with the method of HE stain, and used the immunohistochemistry to investigate the expression of BMP-2 and TGF-β1 in bone tissues.Result:Is The comparison of the BMD of the femur of the rats in each experimentation groups:Comparing with the sham operation group, the BMD of the femur of the model group has evident tendeney to reduce; comparing with the model group, in the nomal dose herb group of Bushen Hutan Fang,the much dose herb group of Bushen Hutan Fang and the western medicine group, the BMD of femur increased evidently.2,The comparison of the blood biochemisty index of the experimental rats:Comparing with the sham operation group, the activity of N-MID in blood serum of model group increased evidently; comparing with the model group, the activity of N-MID in blood serum of the nomal dose herb group of Bushen Hutan Fang,the much dose herb group of Bushen Hutan Fang and the western medicine group decreased evidently; Comparing with the sham operation group, the level of 25 (OH) VD and PICP in blood serum of the rat of model group decreased remarkably, comparing with the model group, the activity of 25 (OH) VD and PICP in blood serum of the nomal dose herb group of Bushen Hutan Fang, the much dose herb group of Bushen Hutan Fang and the western medicine group increased evidently.3,RT-PCR detection:the level of the expression of the gene of BGPmRNA and CbfalmRNA in bone marrow cells of the model group decreased evidently, the treatment of the nomal dose herb group of Bushen Hutan Fang,the much dose herb group of Bushen Hutan Fang and the western medicine group can inerease the mRNA expression.4,The comparison of the morpholoy of the bone tissues in the experimental rats:we observed the HE stain right tibia of the rats in the LM:in sham operation group, the arrangement of the bone trabeeula are normal complete, the shape and strueture are complete; In the model group, the arrangement of bone trabecula strueture is rarefaction, it appeared, he bone trabecula thickness decreased, many caecum of the bone trabeeula, the marrow cavity area rates inreased obviously.5. Immunohistochemistry method to detect the BMP-2 and TGF-β1 in the bone tissues:in sham operation group,, the BMP-2 and TGF-β1 expression site is the bone matrix and the osteoblast surrounding the bone trabecula. The main place is the cytoplasm of the osteoblast; Comparing with the sham operation group, the level of the expression of the BMP-2 and TGF-β1 in bone tissues of the model group decreased evidently, the treatment of the nomal dose herb group of Bushen Hutan Fang,the much dose herb group of Bushen Hutan Fang and the western medicine group can inerease the BMP-2 and TGF-β1 expression.Conclusion:The experiment result indicate:The Bushen Hutan Fang can enhance the BMD of the experimental rats, increase the content of 25 (OH) VD and PICP in blood serum, decrease the content of N-MID in blood serum, increase the expression of BGPmRNA and CbfalmRNA in bone marrow cells, increase the expression of BMP-2 and TGF-β1 in bone tissues.The experiment verifies that Bushen Hutan Fang can modulate rat's bone marrow microenvironment, enhance the production of cytokines which profit bone formation, then to effects the treatment of osteoporosis.